The present invention relates generally to the detection and treatment of Chiamydial infection. In particular, the invention is related to polypeptides comprising a Chlamydia antigen and the use of such polypeptides for the serodiagnosis and treatment of Chlamydial infection.
Chlamydiae are intracellular bacterial pathogens that are responsible for a wide variety of important human and animal infections. Chlamydia trachomatis is one of the most common causes of sexually transmitted diseases and can lead to pelvic inflammatory disease (PID), resulting in tubal obstruction and infertility. Chlamydia trachomatis may also play a role in male infertility. In 1990, the cost of treating PID in the US was estimated to be $4 billion. Trachoma, due to ocular infection with Chlamydia trachomatis, is the leading cause of preventable blindness worldwide. Chlamydia pneumonia is a major cause of acute respiratory tract infections in humans and is also believed to play a role in the pathogenesis of atherosclerosis and, in particular, coronary heart disease. Individuals with a high titer of antibodies to Chlamydia pneumonia have been shown to be at least twice as likely to suffer from coronary heart disease as seronegative individuals. Chlamydial infections thus constitute a significant health problem both in the US and worldwide.
Chlamydial infection is often asymptomatic. For example, by the time a woman seeks medical attention for PID, irreversible damage may have already occurred resulting in infertility. There thus remains a need in the art for improved vaccines and pharmaceutical compositions for the prevention and treatment of Chlamydia infections. The present invention fulfills this need and further provides other related advantages.
The present invention provides compositions and methods for the diagnosis and treatment of Chlamydia infection. In one aspect, polypeptides are. provided comprising an immunogenic portion of a Chlamydia antigen, or a variant of such an antigen. In one embodiment, the antigen comprises an amino acid sequence encoded by a DNA sequence selected from the group consisting of (a) a sequence of SEQ ID NO: 1, 15 or 21-25; (b) the complements of said sequences; and (c) sequences that hybridize to a sequence of (a) or (b) under moderately stringent conditions. In a specific embodiment, a polypeptide comprising an amino acid sequence of SEQ ID NO: 5 is provided.
In a related aspect, polynucleotide sequences encoding the above polypeptides, recombinant expression vectors comprising one or more of these polynucleotide sequences and host cells transformed or transfected with such expression vectors are also provided.
In another aspect, the present invention provides fusion proteins comprising an inventive polypeptide, or, alternatively, an inventive polypeptide and a known Chlamydia antigen. In yet another aspect, the present invention provides antibodies, both polyclonal and monoclonal, that bind to the polypeptides described. above.
Within other aspects, the present invention provides pharmaceutical compositions that comprise one or more Chlamydia polypeptides disclosed herein, or a polynucleotide molecule encoding such polypeptides, and a physiologically acceptable carrier. The invention also provides vaccines comprising one or more of the disclosed polypeptides and a non-specific immune response enhancer, together with vaccines comprising one or more polynucleotide sequences encoding such polypeptides and a non-specific immune response enhancer.
In yet another aspect, methods are provided for inducing protective immunity in a patient, comprising administering to a patient an effective amount of one or more of the above pharmaceutical compositions or vaccines.
In yet a further aspect, methods for the treatment of Chlamydia infection in a patient are provided, the methods comprising obtaining PBMC from the patient, incubating the PBMC with a polypeptide of the present invention (or a polynucleotide that encodes such a polypeptide) to provide incubated T cells and administering the incubated T cells to the patient. The present invention additionally, provides methods for the treatment of Chlamydia infection that comprise incubating antigen presenting cells with a polypeptide of the present invention (or a polynucleotide that encodes such a polypeptide) to provide incubated antigen presenting cells and administering the incubated antigen presenting cells to the patient. In certain embodiments, the antigen presenting cells are selected from the group consisting of dendritic cells, macrophages, monocytes, B-cells, and fibroblasts. Compositions for the treatment of Chlamydia infection comprising T cells or antigen presenting cells that have been incubated with a polypeptide or polynucleotide of the present invention are also provided.
In further aspects of the subject invention, methods and diagnostic kits are provided for detecting Chlamydia infection in a patient. In one embodiment, the method comprises: (a) contacting a biological sample with at least one of the polypeptides or fusion proteins disclosed herein; and (b) detecting in the sample the presence of antibodies that bind to the polypeptide or fusion protein, thereby detecting Chlamydia infection in the biological sample. Suitable biological samples include whole blood, sputum, serum, plasma, saliva, cerebrospinal fluid and urine. In one embodiment, the diagnostic kits comprise one or more of the polypeptides or fusion proteins disclosed herein in combination with a detection reagent. In yet another embodiment, the diagnostic kits comprise either a monoclonal antibody or a polyclonal antibody that binds with a polypeptide of the present invention.
The present invention also provides methods for detecting Chlamydia infection comprising: (a) obtaining a biological sample from a patient; (b) contacting the sample with at least two oligonucleotide primers in a polymerase chain reaction, at least one of the oligonucleotide primers being specific for a polynucleotide sequence disclosed herein; and (c) detecting in the sample a polynucleotide sequence that amplifies in the presence of the oligonucleotide primers. In one embodiment, the oligonucleotide primer comprises at least about 10 contiguous nucleotides of a polynucleotide sequence eptide disclosed herein, or of a sequence that hybridizes thereto.
In a further aspect, the present invention provides a method for detecting Chlamydia infection in a patient comprising: (a) obtaining a biological sample from the patient; (b) contacting the sample with an oligonucleotide probe specific for a polynucleotide sequence disclosed herein; and (c) detecting in the sample a polynucleotide sequence that hybridizes to the oligonucleotide probe. In one embodiment, the oligonucleotide probe comprises at least about 15 contiguous nucleotides of a polynucleotide sequence disclosed herein, or a sequence that hybridizes thereto.
These and other aspects of the present invention will become apparent upon reference to the following detailed description. All references disclosed herein are hereby incorporated by reference in their entirety as if each was incorporated individually.
SEQ ID NO: 1 is the determined DNA sequence for the C. trachomatis clone 1-B1-66.
SEQ ID NO: 2 is the determined DNA sequence for the C. trachomatis clone 4-D7-28.
SEQ ID NO: 3 is the determined DNA sequence for the C. trachomatis clone 3-G3-10.
SEQ ID NO: 4 is the determined DNA sequence for the C. trachomatis clone 10-C10-31.
SEQ ID NO: 5 is the predicted amino acid sequence for 1-B1-66.
SEQ ID NO: 6 is the predicted amino acid sequence for 4-D7-28.
SEQ ID NO: 7 is a first predicted amino acid sequence for 3-G3-10.
SEQ ID NO: 8 is a second predicted amino acid sequence for 3-G3-10.
SEQ ID NO: 9 is a third predicted amino acid sequence for 3-G3-10.
SEQ ID NO: 10 is a fourth predicted amino acid sequence for 3-G3-10.
SEQ ID NO:11 is a fifth predicted amino acid sequence for 3-G3-10.
SEQ ID NO: 12 is the predicted amino acid sequence for 10-C10-31.
SEQ ID NO: 13 is the amino acid sequence of the synthetic peptide 1-B1-66/49-67.
SEQ ID NO: 14 is the amino acid sequence of the synthetic peptide 1-B1-66/58-77.
SEQ ID NO: 15 is the determined DNA sequence for the C. trachomatis serovar LGV II clone 2C7-8.
SEQ ID NO: 16 is the determined DNA sequence for a first putative open reading frame from C. trachomatis serovar D.
SEQ ID NO: 17 is the predicted amino acid sequence encoded by the first putative open reading frame from C. trachomatis serovar D.
SEQ ID NO: 18 is the determined amino acid sequence of the synthetic peptide CtC7.8-12.
SEQ ID NO: 19 is the determined amino acid sequence of the synthetic peptide CtC7.8-13.
SEQ ID NO: 20 is the predicted amino acid sequence encoded by a second putative open reading from C. trachomatis serovar D.
SEQ ID NO: 21 is the determined DNA sequence for clone 4C9-18 from C. trachomatis LGV II.
SEQ ID NO: 22 is the determined DNA sequence homologous to Lipoamide Dehydrogenase from C. trachomatis LGV II.
SEQ ID NO: 23 is the determined DNA sequence homologous to Hypothetical protein from C. trachomatis LGV II.
SEQ ID NO: 24 is the determined DNA sequence homologous to Ubiquinone Mehtyltransferase from C. trachomatis LGV II.
SEQ ID NO: 25 is the determined DNA sequence for clone 4C9-18#2 BL21 pLysS from C. trachomatis LGV II.
SEQ ID NO: 26 is the predicted amino acid sequence for 4C9-18#2 from C. trachomatis LGV II. 
SEQ ID NO: 27 is the determined DNA sequence for Cp-SWIB from C. pneumonia strain TWAR.
SEQ ID NO: 28 is the predicted amino acid sequence for Cp-SWIB from C. pneumonia strain TWAR.
SEQ ID NO: 29 is the determined DNA sequence for Cp-S13 from C. pneumonia strain TWAR.
SEQ ID NO: 30 is the predicted amino acid sequence for Cp-S13 from C. pneumonia strain TWAR.
SEQ ID NO: 31 is the amino acid sequence for a 10 mer consensus peptide from CtC7.8-12 and CtC7.8-13.
SEQ ID NO: 32 is the predicted amino acid sequence for clone 2C7-8 from C. trachomatis LGV II.
SEQ ID NO: 33 is the determined DNA sequence of a clone from C. trachomatis serovar D which shows homology to clone 2C7-8.
SEQ ID NO: 34 is the predicted amino acid sequence encoded by the sequence of SEQ ID NO: 33.
SEQ ID NO: 35 is the determined DNA sequence for C.p. SWIB Nde (5xe2x80x2 primer) from C. pneumonia. 
SEQ ID NO: 36 is the determined DNA sequence for C.p. SWIB EcoRI (3xe2x80x2 primer) from C. pneumonia. 
SEQ ID NO: 37 is the determined DNA sequence for C.p. S13 Nde (5xe2x80x2 primer) from C. pneumonia. 
SEQ ID NO: 38 is the determined DNA sequence for C.p. S13 EcoRI (3xe2x80x2 primer) C. pneumonia.